| 초록 |
In this study, cloning of cold-active lipase from psychrophile Janthinobacterium sp. was carried out. From the partial genome sequencing of Janthinobacterium sp. PAMC 25641, putative gene encoding lipase was cloned into a pET28a(+) vector and heterologously expressed in Escherichia coli BL21(DE3). The amino acid sequence deduced from the nucleotide sequence (930 bp) corresponded to a protein of 309 amino acid residues with a molecular weight of 32.7 kDa and a pI of 5.55. The deduced protein showed 93% and 92% amino acid sequence identities with the lipases from Janthinobacterium lividum and Janthinobacterium sp. HH107, respectively. The recombinant E.coli cells harboring putative lipase of Janthinobacterium sp. were induced by addition of isopropyl-β-D-thiogalactopyranoside (IPTG). To purify lipase, inclusion body was solubilized and Ni2+-NTA affinity chromatography was carried. |
